Background of CDC25A antibody

The cdc25 protein phosphatase family plays a critical role in activating cyclin-dependent kises (CDKs) via dephosphorylation of conserved Thr14/Tyr15 inhibitory phosphorylation sites. While cdc25C is primarily responsible for activating CDK1 to overcome the G2/M checkpoint and allow mitotic entry, the primary substrate of cdc25A is CDK2, which, when active, allows progression through the G1/S and intra-S checkpoints. Abundance, subcellular localization and activity of cdc25A is tightly controlled by a variety of mechanisms, including phosphorylation, ubiquitition, and inhibitory binding to 14-3-3 proteins. During normal cell cycle progression, elevated c-Myc and E2F transcription factor levels lead to increased cdc25A expression. When conditions are favorable for D synthesis, cdc25A and CDK2 form an activation loop, wherein each activates the other enzyme. D damage, on the other hand, leads to multisite phosphorylation at inhibitory sites (Ser123, Ser177, Ser278, Ser292, and Thr506) by Chk1 and Chk2, which result in 14-3-3 binding and ubiquitin-mediated degradation.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with CDC25A (phospho S124) polyclonal antibody ( Cat # PAB0425 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. HC = hepatocarcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with CDC25A (phospho S278) polyclonal antibody ( Cat # PAB0427 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with CDC25A (phospho S292) polyclonal antibody ( Cat # PAB0428 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. HC = hepatocarcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with CDC25A (phospho S75) polyclonal antibody ( Cat # PAB0429 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with CDC25A (phospho T506) polyclonal antibody ( Cat # PAB0430 ) which was peroxidase-conjugated to the secondary antibody followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

The anti-Phospho-CDC25A pSer292 Pab (Cat. #AP12579PU-N) is used in Western blot to detect Phospho-CDC25A-S292 in cells transfected with wild type or mutant S292A of CDC25A. Data courtesy of Dr. Tiebang Kang of Washington University, St. Louis, MO.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.

The anti-Phospho-CDC25A pThr506 Pab (Cat. #AP12581PU-N) is used in Western blot to detect Phospho-CDC25A-T506 in cells transfected with wild type or mutant T506A of CDC25A. Data courtesy of Dr. Tiebang Kang of Washington University, St. Louis, MO.

Figure 1. Immunoprecipitation of CDC25A from Raji cells with normal Mouse IgG (Lane 1) or AM20026AF-N (Lane 2). After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoblotted with CDC25A antibody (AM20026AF-N).

Figure 3. Immunoprecipitation of CDC25A from Raji cells with Mouse IgG (Lane 1) or AM05219AF-N (Lane 2). After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoblotted with CDC25A antibody (AM05219AF-N).

Immunohistochemical analysis of CDC25A (pS75) staining in human tonsil formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CDC25A (RC214325, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CDC25A.

Immunohistochemical analysis of CDC25A (pS178) staining in human prostate cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.